Mitochondrial Protease ClpP is a Target for the Anticancer Compounds ONC201 and Related Analogues

ONC201 is a first-in-class imipridone molecule currently in clinical trials for the treatment of multiple cancers. Despite enormous clinical potential, the mechanism of action is controversial. To investigate the mechanism of ONC201 and identify compounds with improved potency, we tested a series of novel ONC201 analogues (TR compounds) for effects on cell viability and stress responses in breast and other cancer models. The TR compounds were found to be ∼50-100 times more potent at inhibiting cell proliferation and inducing the integrated stress response protein ATF4 than ONC201. Using immobilized TR compounds, we identified the human mitochondrial caseinolytic protease P (ClpP) as a specific binding protein by mass spectrometry. Affinity chromatography/drug competition assays showed that the TR compounds bound ClpP with ∼10-fold higher affinity compared to ONC201. Importantly, we found that the peptidase activity of recombinant ClpP was strongly activated by ONC201 and the TR compounds in a dose- and time-dependent manner with the TR compounds displaying a ∼10-100 fold increase in potency over ONC201. Finally, siRNA knockdown of ClpP in SUM159 cells reduced the response to ONC201 and the TR compounds, including induction of CHOP, loss of the mitochondrial proteins (TFAM, TUFM), and the cytostatic effects of these compounds. Thus, we report that ClpP directly binds ONC201 and the related TR compounds and is an important biological target for this class of molecules. Moreover, these studies provide, for the first time, a biochemical basis for the difference in efficacy between ONC201 and the TR compounds

Informing Intercultural Dress Codes

The purpose of this project was to address and understand the present gaps in dress code policy at the University of British Columbia (UBC) Recreation fitness facilities. An objective that corresponds with this purpose was to identify key dress code components that promote interculturalism and inclusion. Additionally, research aimed to explore underlying clothing barriers that hinder an individual’s comfort level when entering a fitness facility. The methods used to gather information included a literature review and a survey developed using the Qualtrics software. Ideally, the findings from this paper will lead to the development of a culturally sensitive dress code that acknowledges intercultural accessibility and inclusion. An in-depth literature review highlighted a number of interesting trends. The majority of Canadian universities had a dress code present, however student and staff opinions on dress code implementation were mixed (BCIT, 2019; Lopatka, 2018). It was also found that employees of private gyms often lacked proper education and knowledge regarding dress code standards at their facility. Finally, literature showed that females can often feel targeted and may face a double standard when dress codes are inconsiderate of gender equality (Graf, 2019). Overall, public perceptions on the effectiveness of dress codes appears to be varied. Based on survey results that examined community member opinions on dress code policy, a number of insights were gathered. The majority of participants denied the idea that dress code policy impacts their decision to attend a specific facility (refer to Appendix C, Figure 1). Similarly, many respondents valued the autonomy to wear what they please and did not want their clothing choices restricted (refer to Appendix C, Figure 2). However, results were conflicting as the majority of participants responded that they would be willing to comply with a dress code if it was implemented to ensure the comfort of all patrons (refer to Appendix C, Figure 3). Men not wearing shirts were most commonly cited as making respondents uncomfortable. However, laced clothing and exposed midriffs also made participants uneasy, but to a lesser degree (refer to Appendix C, Table 1) . At the conclusion of the literature review and survey, five key recommendations were made. The first recommendation is to improve employee education regarding dress codes so that all employees are well versed and consistent with dress code knowledge. Furthermore, dress codes must utilize clear and concise language to limit ambiguity for gym attendees. To support this recommendation, specific rules posted in numerous accessible formats should be developed. Another recommendation is to ensure gender equality by holding all genders to the same standard in regards to what body parts they can expose. Additionally, promoting gym dress codes from a stance of health and hygiene may aid in limiting negative responses by patrons who may otherwise feel targeted. Finally, it was clear that dress codes should not impact cultural and religious clothing. Instead, cut off tee shirts, clothing that is laced and clothing that exposes the midriff should be considered by fitness centre management when creating a dress code. Disclaimer: “UBC SEEDS provides students with the opportunity to share the findings of their studies, as well as their opinions, conclusions and recommendations with the UBC community. The reader should bear in mind that this is a student project/report and is not an official document of UBC. Furthermore readers should bear in mind that these reports may not reflect the current status of activities at UBC. We urge you to contact the research persons mentioned in a report or the SEEDS Coordinator about the current status of the subject matter of a project/report.”Education, Faculty ofKinesiology, School ofUnreviewedUndergraduat